The response of cells to mechanical inputs is a key determinant of cell behavior. In response to changes in the mechanical environment of epithelial cells, E-cadherin initiates signal transduction cascades that allow the cells to modulate their contractility to withstand the force. Much attention has focused on identifying the E-cadherin signaling pathways that promote contractility, but the negative regulators remain undefined. In this thesis, we identify SHP2 as a force-activated phosphatase that negatively regulates E-cadherin force transmission by dephosphorylating vinculin Y822. To specifically probe a role for SHP2 in E-cadherin mechanotransduction, we innovatively mutated vinculin so that it retains its phosphorylation but cannot be dephosphorylated. Cells expressing the mutant vinculins have increased contractility. This work provides the first mechanism for inactivating E-cadherin mechanotransduction and provides a new method for specifically targeting the action of phosphatases in cells.

Glass Fibre-Reinforced Polymer (GFRP) bars offer a feasible alternative in locations where steel is not the suitable reinforcement; namely locations that are sensitive to corrosion. In this study 60 specimens, 52 GFRP reinforced and 8 steel reinforced, were constructed and tested under direct tension in order to investigate the tension stiffening and cracking behaviour. The effects of different variables such as the bar type, the bar diameter, the reinforcement ratio and the concrete strength on tension stiffening and crack spacing were studied. The current code provisions for tension stiffening, namely ACI-440 and CEB-FIP were evaluated against the obtained test data. It was determined that the current code provisions significantly overestimate tension stiffening in GFRP reinforced specimens. A new tension stiffening model was, therefore, developed that provides better simulation of the test data. The CEB-FIP 1978 model for crack spacing was also modified for GFRP reinforced members.

Glass Fibre-Reinforced Polymer (GFRP) bars offer a feasible alternative in locations where steel is not the suitable reinforcement; namely locations that are sensitive to corrosion. In this study 60 specimens, 52 GFRP reinforced and 8 steel reinforced, were constructed and tested under direct tension in order to investigate the tension stiffening and cracking behaviour. The effects of different variables such as the bar type, the bar diameter, the reinforcement ratio and the concrete strength on tension stiffening and crack spacing were studied. The current code provisions for tension stiffening, namely ACI-440 and CEB-FIP were evaluated against the obtained test data. It was determined that the current code provisions significantly overestimate tension stiffening in GFRP reinforced specimens. A new tension stiffening model was, therefore, developed that provides better simulation of the test data. The CEB-FIP 1978 model for crack spacing was also modified for GFRP reinforced members.

No description available.

The rolling of an aircraft about its fuselage produces centrifugal forces which affect the stiffness of the wings. A number of previous studies explain the effect of centrifugal stiffening in rotating beams and consequently on the frequencies of the beam. Multiple cases of the rotating beam are explored in this thesis to investigate effects of mass distribution and boundary conditions on the frequencies of centrifugally stiffened beams. It is found that for a uniform beam with all degrees of freedom free on both ends, the rigid modes of the beam are affected and are no longer zero when it is stiffened from centrifugal forces. This thesis aims to set up a model to investigate the stiffening effects using the mAEWing2 aircraft. A preliminary analysis is done for the mAEWing2 aircraft and the roll rate, control surface deflection and angle of attack are identified as the parameters to be studied. For a given angle of attack and control surface deflection, the centrifugal forces in the aircraft in steady roll are determined using trim analysis. These are used to pre-stress the model for modal analysis. It is found that in mAEWing2 aircraft in steady roll maneuvers, the centrifugal stiffening effect on the natural frequencies is not significant. It emphasizes the need to conduct a sensitivity analysis to include centrifugal stiffening in the dynamic analysis while designing an aircraft. This, along with some de-stiffening due to gravity loads might be important for the future N+3 aircraft with their high aspect ratio large wingspans. / MS / Structural analysis is mainly concerned with determining the behavior of a structure when subjected to a disturbance. The natural response of a structure to some disturbance is termed as free vibration of the structure. The term vibration describes repetitive motion that can be observed in a structure and is influenced by its material and structural properties. These vibrations may cause fatigue in the structure and the performance of the structure may be adversely affected. Consequently it becomes necessary to study and eliminate these vibrations. The vibration characteristics of a system are described by its natural frequencies and mode shapes. Natural frequencies of a structure are the frequencies at which the structure naturally tends to vibrate if it is subjected to a disturbance. The deformed shape of the structure vibrating at one of its specific natural frequencies of vibration is termed its normal mode shape. In the case of a rotating beam, the centrifugal force acts axially along the length of the beam. When the rotating beam deflects upwards, the centrifugal force creates a downward bending moment, reducing its net deflection. The ratio of force to displacement increases, increasing the stiffness of the rotating beam. This effect is called the stiffening effect. There is a large volume of literature that presents the effect of stiffening on the natural frequencies of a rotating beam model, for various boundary conditions. Such a stiffening analysis has also been done for the blades of a turbine and turbo fans. In addition, there are models available for analyzing the aerodynamic model of an aircraft in roll, considering stability derivatives of the aircraft. However, there are gaps in the available literature in analyzing an aircraft in roll from the perspective of structural analysis. The rolling of an aircraft about its fuselage produces centrifugal forces which affect the stiffness of the wings. A number of previous studies explain the effect of centrifugal stiffening in rotating beams and consequently on the frequencies of the beam. Multiple cases of the rotating beam are explored in this thesis to investigate effects of mass distribution and boundary conditions on the frequencies of centrifugally stiffened beams. It is found that for a uniform beam with all degrees of freedom free on both ends, the rigid modes of the beam are affected and are no longer zero when it is stiffened from centrifugal forces. This further motivates the need for investigating the effect of centrifugal stiffening in spinning spacecraft and aircraft in rapid roll maneuvers. This thesis further aims to set up a model to investigate the stiffening effects using the mAEWing2 aircraft. A preliminary analysis is done for the mAEWing2 aircraft and the roll rate, control surface deflection and angle of attack are identified as the parameters to be studied. For a given angle of attack and control surface deflection, the centrifugal forces in the aircraft in steady roll are determined using trim analysis. These are used to pre-stress the model for modal analysis. It is found that in the mAEWing2 aircraft in steady roll maneuvers the stiffening effect on the frequencies is not significant. It emphasizes the need to conduct a sensitivity analysis to include centrifugal stiffening in the dynamic analysis while designing an aircraft. This, along with some de-stiffening due to gravity loads might be important for the future N+3 aircraft with their high aspect ratio large wingspans.

The objective of the present study was to; 1) determine if the use of a continuous cognitive task demonstrates distinct characteristics of a more automatic control of posture, compared to an external focus (EF) and feet together (FT) postural task, and to 2) examine which condition, if any, exhibits the characteristics of increased ankle stiffness proposed by Winter et al. (1998), as well as displaying increased ankle muscular co-contractions, which are a suggested neuromuscular mechanism that stiffens posture. Fifteen young adults stood on a force platform and performed 4 separate conditions: FT, EF, single number sequence (SNS), and double number sequence (DNS). Throughout the session, surface electromyography (EMG) signals were collected from the tibialis anterior (TA) and medial gastrocnemius (MG) of each leg. Each testing session consisted of 24 trials, with 6 per condition. Results displayed decreased sway area for SNS and DNS compared to FT. Sway variability in the anterior/posterior (AP) direction SNS and DNS were smaller compared to EF and FT. As well sway variability in the medial/lateral (ML) direction was smaller for SNS and DNS compared to FT. ML Mean velocity (MV) did not differ across conditions, though in the AP direction it was larger for SNS and DNS compared to EF and FT. AP Mean power frequency (MPF) was larger for SNS compared to FT. In the ML direction MPF was larger for SNS and DNS compared to FT. Co-Contraction indices revealed no differences across conditions. Conversely the left TA for DNS revealed increased EMG activation compared to EF and SNS.

No description available.

Gels and hydrogels have attracted a great attention for potential applications in tissue engineering, drug delivery, actuators, and soft robots. There has been a significant progress to engineer hydrogels from both synthetic and natural precursors to be as tough as a solid and as stretchable as a rubbery material while maintaining high water/solvent content. Despite considerable advances in rationally designing hydrogels, our understanding of their complex nonlinear mechanical deformation behavior is incomplete. This is partially due to the difficulty in conducting mechanical characterization on slippery, soft and swollen gels. Thus, it is required to develop new experimental techniques in order to better characterize them. Further, analyzing the experimental observations and link it with the molecular networks is an important factor. With this perspective, in this dissertation, nonlinear mechanical properties of different gel like materials have been investigated. We chose different gels with varied molecular structure, from molecular gel to self-assembled copolymer gels with flexible chains, to semiflexible polysaccharide based polymers. By developing suitable experimental protocols, strain-stiffening behavior of these materials, similar to that observed in biological materials, have been captured. Chain flexibility is a dominant factor in mechanical behavior of gels. For example, gels with flexible chains dilate orthogonal to an external shear load, whereas gels with semilexible chains contract similar to biological gel-like materials. In order to investigate the failure mechanism in our gels, cavitation rheology technique was also applied. We found that cavitation phenomenon in gels is related to the molecular architecture of the gels. The present work provides a better understanding of the deformation behavior of soft gels when subjected to a large load.

Background: Vascular stiffening develops with both hypertension and aging, and is a strong predictor of end-organ damage. Excessive deposition of collagen by vascular smooth muscle cells (VSMCs) can lead to decreased compliance of vessels such as the aorta. The IRE1α arm of the unfolded protein response is activated in cells with a secretory phenotype due to its role in augmenting protein folding capacity. We hypothesize that by a similar mechanism, VSMCs transitioning to a collagen-secreting phenotype in response to TGF-β1 require the activation of IRE1. Inhibition of this pathway is hypothesized to reduce collagen secretion and hence prevent the development of fibrosis in the aorta. Methods: Collagen deposition by VSMCs in vitro was measured using immunoblotting and a Picrosirius Red-based colorimetric assay. Western blot and qRT-PCR were used to assess the expression of ER stress markers. Ex vivo culture of aortic rings was also performed to determine the effect of 4µ8c on TGF-β1-induced vascular stiffening. 12-14 week old male spontaneously hypertensive rats were divided into three treatment groups: 1) No treatment, 2) L-NAME (50 mg/L), and 3) L-NAME and the IRE1α inhibitor 4µ8c (2.5 mg/kg/day i.p.). Aortic compliance after 18 days of treatment was measured ex vivo using a wire myograph to construct tension-diameter curves. Results: Inhibition of IRE1α endonuclease activity by 4µ8c reduced collagen production in VSMCs stimulated with TGF-β1 or Ang II. A decrease in the expression of the collagen-associated chaperones PDI, GRP78 and GRP94 was observed. Aortic rings treated with TGF-β1 developed vascular stiffening, which was improved by co-treatment with 4µ8c. SHRs treated with L-NAME for 18 days developed aortic stiffening, which was prevented by daily injections of 4µ8c. Conclusions: Our data suggest that inhibition of the IRE1α pathway can reduce vascular stiffening and fibrosis by disrupting the collagen biosynthesis pathway in VSMCs. / Thesis / Master of Science (MSc)

Cells respond to their mechanical environment by changing shape and size, migrating, or even differentiating to a more specialized cell type. A better understanding of the response of cells to surrounding cues will allow for more targeted and effected designs for biomedical applications, such as disease treatment or cellular therapy. The spreading behavior of both human mesenchymal stem cells (hMSCs) and 3T3 fibroblasts is a function of substrate stiffness, and can be quantified to describe the most visible response to how a cell senses stiffness. The stiffness of the substrate material can be modulated by altering the substrate thickness, and this has been done with the commonly-used linearly elastic gel, polyacrylamide (PA). Though easy to produce and tune, PA gel does not exhibit strain-stiffening behavior, and thus is not as representative of biological tissue as fibrin or collagen gel. Fibroblasts on soft fibrin gel show spreading similar to much stiffer linear gels, indicating a difference in cell stiffness sensing on these two materials. We hypothesize cells can sense further into fibrin and collagen gels than linear materials due to the strain-stiffening material property. The goal of this work is to compare the material response of linear (PA) and strain-stiffening (fibrin, collagen gel) substrates through modulation of effective stiffness of the materials. The two-step approach is to first develop a finite element model to numerically simulate a cell contracting on substrates of different thicknesses, and then to validate the numerical model by quantifying fibroblast spreading on sloped fibrin and collagen gels. The finite element model shows that the effective stiffness of both linear and nonlinear materials sharply increases once the thickness is reduced below 10µm. Due to the strain-stiffening behavior, the nonlinear material experiences a more drastic increase in effective stiffness at these low thicknesses. Experimentally, the gradual response of cell area of HLF and 3T3 fibroblasts on fibrin and collagen gels is significantly different (p<0.05) from these cell types on PA gel. This gradual increase in area as substrate thickness decreases was not predicted by the finite element model. Therefore, cell spreading response to stiffness is not explained by just the nonlinearity of the material.